Menu

Javascript is not activated in your browser. This website needs javascript activated to work properly.
You are here

Epitope analysis of insulin autoantibodies using recombinant Fab

Author:
  • C J Padoa
  • N J Crowther
  • J W Thomas
  • T R Hall
  • L M Bekris
  • Carina Törn
  • Mona Landin-Olsson
  • E Ortqvist
  • J P Palmer
  • Åke Lernmark
  • C S Hampe
Publishing year: 2005
Language: English
Pages: 564-571
Publication/Series: Clinical and Experimental Immunology
Volume: 140
Issue: 3
Document type: Journal article
Publisher: British Society for Immunology

Abstract english

Autoantibodies to insulin are often the first autoantibodies detected in young children with type 1 diabetes and can be present before the onset of clinical diabetes. These autoantibodies and their epitopes are, however, not well characterized. We explored the use of monoclonal antibodies and their recombinant Fab as reagents for epitope analysis. In this study we cloned and characterized the recombinant Fab of the insulin-specific monoclonal antibody CG7C7. We found the epitope of this antibody to be located predominantly at the A-chain loop of the insulin molecule. The recombinant Fab was then used to compete for insulin binding against insulin autoantibodies present in sera from patients with type 1 or type 1.5 diabetes. In competition experiments with sera positive for autoantibodies to insulin the recombinant Fab significantly reduced the binding to [I-125]-insulin by sera of type 1 (n = 35) and type 1.5 diabetes [latent autoimmune diabetes in adults (LADA)] (n = 14) patients (P < 0.0001). We conclude that competition between insulin-specific monoclonal antibodies or their recombinant Fab and insulin autoantibodies should prove useful in the epitope analysis of autoantibodies to insulin.

Keywords

  • Immunology in the medical area
  • type 1 diabetes
  • epitopes
  • insulin autoantibody
  • radioligand binding
  • assay
  • recombinant Fab

Other

Published
  • Diabetes and Celiac Unit
  • ISSN: 0009-9104
E-mail: ake [dot] lernmark [at] med [dot] lu [dot] se

Principal investigator

Diabetes and Celiac Unit

+46 40 39 19 01

+46 70 616 47 79

60:11:015

Jan Waldenströms gata 35, Malmö

33

Lund University Diabetes Centre, CRC, SUS Malmö, Jan Waldenströms gata 35, House 91:12. SE-214 28 Malmö. Telephone: +46 40 39 10 00